DEVELOPMENT AND EVALUATION OF NEW SCREENING TESTS

本提案的目标和具体目的是， 用于检测针对HTLV-I、HTLV-II和HTLV-III的抗体的测定 HIV-2抗原以及检测、表征和 这些相同病毒的蛋白质和核酸的定量。 所有试验将利用先前累积和充分表征的 超过20，000人血清和细胞库，以便于评估 灵敏度和特异性。 将设计血清学试验检测IgG或IgM 反应性测定形式将涉及多种病毒来源， 抗原包括纯化的病毒粒子、合成肽和重组 proteins.目的将是开发ELISA、蛋白质印迹和/或免疫印迹。 免疫印迹足够敏感和特异， 为RIPA的。 蛋白质检测将集中在多克隆和 用于直接细胞染色的单克隆抗体或抗原 捕获测定格式。抗体将针对所有主要的 结构、功能和调节病毒蛋白。 核酸研究将集中在PCR的继续发展上 和QB复制酶系统进行检测、定量和测序 从人体组织和体液中提取特定的病毒DNA和RNA。 预计上述技术将进入 预防、诊断和治疗人类疾病的临床系统 逆转录病毒疾病

The objectives and specific aims of this proposal are to develop improved assays for both the detection of antibodies against HTLV-I, HTLV-II and HIV-2 antigens and also the detection, characterization, and quantification of the proteins and nucleic acids of these same viruses. All assays will make use of previously accrued and well-characterized bank of over 20,000 human sera and cells to facilitate assessment of sensitivity and specificity. Serological assays will be designed to detect either IgG or IgM reactivities. Assay formats will involve a variety of sources of viral antigen including purified virions, synthetic peptides and recombinant proteins. The aim will be to develop ELISA, western blots and/or immunoblots that are sensitive and specific enough to obviate the need for RIPA's. Protein detection will center on the development of polyclonal and monoclonal antibodies to be used in direct cell staining or antigen capture assay formats. Antibodies will be made against all major structural, functional, and regulatory viral proteins. Nucleic acid studies will center on the continued development of the PCR and QB replicase system for the detection, quantification and sequencing of specific viral DNA and RNA from human tissues and body fluids. It is anticipated that the above techniques will make their way into clinical systems for the prevention, diagnosis and treatment of human retroviral disorders.